Development of a Competitive Time-Resolved Fluoroimmunoassay for Paclitaxel.

INTRODUCTION: Paclitaxel (Tax) is a diterpene alkaloid isolated from Taxus species and has proved clinically effective in treating a number of malignancies. Current quantitative analytical methods for Tax such as high-performance liquid chromatography (HPLC) often involve complicated sample preparation procedures with low recovery rates. OBJECTIVE: To establish a rapid and sensitive time-resolved fluoroimmunoassay (TRFIA) for measuring Tax in Taxus materials with convenient sample preparation and a high recovery rate. METHODS: Rabbit anti-mouse IgG was coated onto a 96-well microplate, which was then incubated with standard solutions of Tax and anti-Tax monoclonal antibody 3A3. A Eu3+ -labelled conjugate of Tax and human serum albumin was used as the tracer. The luminescent system was enhanced with a solution containing 2-naphthoyltrifluoroacetone. RESULTS: The established TRFIA showed a linear response within the Tax concentration range of 3.2 to 80 ng/mL, with a limit of detection of 1.4 ng/mL. The intra- and inter-assay coefficient of variation of the assay was 9.6% and 9.7%, respectively, with an average recovery rate from spiked samples of 108.5%. Tax contents in Taxus samples were determined using both the established TRFIA system and a previously established enzyme-linked immunosorbent (ELISA), and the results of two assays were well correlated. CONCLUSION: This TRFIA system shows a high sensitivity, precision and accuracy for detection of Tax. This assay, which is convenient and less time-consuming, allows rapid analysis of Tax and provides another option for Tax measurement for quality control of Taxus materials and products. Copyright © 2017 John Wiley & Sons, Ltd.

[Preliminary study of determination of Shougong in Jinlong Capsules based on fluorescence quantitative PCR method].

A pair of species-specific primer (GZG1/GZG2) based on COⅠ sequence regions for identification of Gekko chinensis were designed. A fluorescent quantitative PCR method was established to identify and quantify G. chinensis from Jinlong Capsules Formula. A standard curve for quantitative analysis of G. chinensis was established (the standard curve equation: y=-3.012 7x+34.501, y is Ct value, x is lg N, N is the copies of COⅠ fragment from G. chinensis). Samples included G. chinensis appeared amplification, while falsify group (not included G. chinensis) and negative control did not have amplification products. The copy number of COⅠ region of G. chinensis was respectively 11.511×106, 6.416×106, 2.553×106 copies/µL in all quality goods, quality goods-adulterants 1:1, quality goods-adulterants 1:4. The results accorded with proportion of adding amount roughly. This study can provide a new strategy for quality control of Chinese patent medicine containing animal drug ingredients.

[Identification of Original Species of Fish Maw by DNA Barcoding].

Objective: To identify the original species of fish maw sold in Guangzhou market by DNA barcoding technology. Methods: Mitochondrial cytochrome C subunit I (CO I) gene fragment of eleven fish maw samples were amplified and sequenced with the self-designed primers. UPGMA phylogenetic tree were constructed for clustering analysis. The species origin of each sample was identified with the identification engine provided in the Barcode of Life Data Systems (BOLD). Results: The self-designed primers were effective in fish maw CO I amplification and sequencing, with success rates both of 100%. BOLD identification and UPGMA clustering analysis indicated the fish maw samples were derived from five fish species of three families. Conclusion: DNA barcoding combined with BOLD identification system can accurately identify the species origin of commercial fish maw.